Vesicles, unilamellar/multilamellar

Liposomes, also known as lipid vesicles, are aqueous compartments enclosed by lipid bilayer membranes [56,57]. Figure 10.11 shows how lipid bilayers are arranged in the liposome and the lipid structures in large unilamellar vesicles and multilamellar vesicles. Lipids consist of two components ... 

Suv, small unilamellare vesicles luv, large unilamellare vesicles mlv, multilamellare vesicles mvv, multivesiculare vesicles (Fig. 4 from [1.34]). 

Fig. 5. Types of liposomes SUV, small unilamellar vesicle LUV, large unilamellar vesicle MLV, multilamellar vesicle MW, multivesicular vesicle...

There are several other ways to entrap solutes inside the liposomes, and the entrapping efficiency depends on the structure of liposomes (small unilamellar, large unilamellar, multilamellar, vesicles, etc.) and from the technique for liposome preparation (Roseman etal., 1978 Cullis etal., 1987 Walde and Ishikawa, 2001). 

At low concentrations, a hollow vesicle results with usually just one double layer and, as the concentration is increased, the number of double layers can increase in a transition from unilamellar vesicles to multilamellar structures. Since the hydro-plilic head groups are exposed on the inside as well as the outside of the vesicular structure this provides an opportunity to entrap hydrophilic guest drug molecules both inside the center of the vesicle and, if multilamellar, between the phospholipid bilayers as well. On the other hand, hydrophobic molecules can become incorporated in the hydrophobic regions of the bilayers where the hydrophobic tails overlap. 

A very suitable method for measurement of the lateral diffusion of molecules adsorbed at the foam film surfaces is Fluorescence Recovery after Photobleaching (FRAP) ([491-496], see also Chapter 2). Measurements of the lateral diffusion in phospholipid microscopic foam films, including black foam films, are of particular interest as they provide an alternative model system for the study of molecular mobility in biological membranes in addition to phospholipid monolayers at the air/water interface, BLMs, single unilamellar vesicles, and multilamellar vesicles. 

Other examples of bilayer structures already mentioned are the sponge phase and bicontinuous cubic phases. The sponge phase has been most studied for nonionic surfactants and is related to common microemulsions. Bilayers may also easily close on themselves to form discrete entities including unilamellar vesicles and multilamellar liposomes. Vesicles are of interest because of the division into inner and outer aqueous domains separated by the bilayer. Vesicles and liposomes are normally not thermodynamically stable (although there are exceptions) and tend to phase separate into a lamellar phase and a dilute aqueous solution. Lipid bilayers are important constituents of living organisms and form membranes, which act as barriers between different compartments. Certain surfactants and lipids may form reversed vesicles, i. e. vesicles with inner and outer oleic domains separated by a (reversed) amphiphile bilayer the bilayer may or may not contain some water. 

Abbreviations SUV, Unilamellar vesicles LUV, Large unilamellar vesicles MLV, Multilamellar vesicles. 

The significance of probe flexibility was examined by Cao, et al, who used QELSS to measure Dp of 32 and 54 nm radius polystyrene spheres, phospho-lipid/cholesterol vesicles, and multilamellar vesicles in aqueous 65 and 1000 kDa polyacrylamides(21). The Dp arose from arelaxation rate that was accurately linear in q -, its concentration dependence was a stretched exponential in c. As seen in Figure 9.12, polyacrylamides are more effective at increasing t] than at reducing Dp, and are more effective at slowing sphere motion than at slowing unilamellar... 

In water, a particle of lecithin exhibits myelin growth, ie, cylindrical sheets that are formed by bdayers and are separated by water which may break up into liposomes (vesicles with a single bilayer of Hpid enclosing an aqueous space). PhosphoHpids more generally form multilamellar vesicles (MLV) (5). These usually are converted to unilamellar vesicles (ULV) upon treatment, eg, sonication. Like other antipolar, surface-active agents, the phosphoHpids are... 

Phospholipids e.g. form spontaneously multilamellar concentric bilayer vesicles73 > if they are suspended e.g. by a mixer in an excess of aqueous solution. In the multilamellar vesicles lipid bilayers are separated by layers of the aqueous medium 74-78) which are involved in stabilizing the liposomes. By sonification they are dispersed to unilamellar liposomes with an outer diameter of 250-300 A and an internal one of 150-200 A. Therefore the aqueous phase within the liposome is separated by a bimolecular lipid layer with a thickness of 50 A. Liposomes are used as models for biological membranes and as drug carriers. 

Liposomes are members of a family of vesicular structures which can vary widely in their physicochemical properties. Basically, a liposome is built of one or more lipid bilayers surrounding an aqueous core. The backbone of the bilayer consists of phospholipids the major phospholipid is usually phosphatidylcholine (PC), a neutral lipid. Size, number of bilayers, bilayer charge, and bilayer rigidity are critical parameters controlling the fate of liposomes in vitro and in vivo. Dependent on the preparation procedure unilamellar or multilamellar vesicles can be produced. The diameter of these vesicles can range from 25 nm up to 50 ym—a 2000-fold size difference. 

Vesicles [10, 11] these aggregates of insoluble natural or artificial amphiphiles in water can have various shapes (spherical, cylindrical). Depending on the preparation conditions, small unilamellar or large multilamellar vesicles can be produced. The structures meet the self-organization criterion, because they are, albeit on a long time scale, dynamic and not in thermodynamic equilibrium, which would in many cases be a macroscopically phase separated lamellar phase. 

Liposomes are formed due to the amphiphilic character of lipids which assemble into bilayers by the force of hydrophobic interaction. Similar assemblies of lipids form microspheres when neutral lipids, such as triglycerides, are dispersed with phospholipids. Liposomes are conventionally classified into three groups by their morphology, i.e., multilamellar vesicle (MLV), small unilamellar vesicle (SUV), and large unilamellar vesicle (LUV). This classification of liposomes is useful when liposomes are used as models for biomembranes. However, when liposomes are used as capsules for drugs, size and homogeneity of the liposomes are more important than the number of lamellars in a liposome. Therefore, "sized" liposomes are preferred. These are prepared by extrusion through a polycarbonate... 

Figure 1. Structure of liposomes and lipid microsphere a), multilamellar vesicle b). unilamellar vesicle c). lipid microsphere. Symbols inside the microsphere indicate di- and tri-acyl glycerol.

The aforementioned results refer to unilamellar membrane models but essentially similar results are obtained in multilamellar vesicles, though the kinetics are more complex in such systems. The numerical values observed in these model membranes simply show that one or more of the aforementioned factors arise however, in the in vivo situation, the preeminent effect is unknown but may well be the proximity of the hydroxyl group to the water interface. 

Figure 10.11 Liposome structures, including multilamellar vesicles (MLV) and large unilamellar...

In model systems for bilayers, one typically considers systems which are composed of one type of phospholipid. In these systems, vesicles very often are observed. The size of vesicles may depend on their preparation history, and can vary from approximately 50 nm (small unilamellar vesicles or SUVs) up to many pm (large unilamellar or LUV). Also one may find multilamellar vesicular structures with more, and often many more than, one bilayer separating the inside from the outside. Indeed, usually it is necessary to follow special recipes to obtain unilamellar vesicles. A systematic way to produce such vesicles is to expose the systems to a series of freeze-thaw cycles [20]. In this process, the vesicles are repeatedly broken into fragments when they are deeply frozen to liquid nitrogen temperatures, but reseal to closed vesicles upon thawing. This procedure helps the equilibration process and, because well-defined vesicles form, it is now believed that such vesicles represent (close to) equilibrium structures. If this is the case then we need to understand the physics of thermodynamically stable vesicles. 

Entrapment of plasmid DNA and/or protein into liposomes entails the preparation of a lipid film from which multilamellar vesicles and, eventually, small unilamellar vesicles (SUVs) are produced. SUVs are then mixed with the plasmid DNA and/or protein destined for entrapment and dehydrated. The dry cake is subsequently broken up and rehydrated to generate multilamellar dehydration-rehydration vesicles (DRV) containing the plasmid DNA and/or protein. On centrifugation, liposome-entrapped vaccines are separated from nonentrapped materials. When required, the DRV are reduced in size by microfluidization in the presence or absence of nonentrapped materials or by employing an alternative method (7) of DRV production, which utilizes sucrose (see below). 

Liposomes consist of many phospholipid bilayers of only few, or just one bilayer (Fig. 5). Therefore multilamellar vesicles (MLV), oligolamellar vesicles (OLV), small unilamellar (SUV), and large unilamellar vesicles (LUV) have to be distinguished. Furthermore, multivesicular liposomes (MVL) may be formed. 

Other pharmaceutical applications have seen the SdFFF applied successfully to monitor droplet size distributions in emulsions, together with their physical state or stability. Some examples are fluorocarbon emulsions, safflower oil emulsions, soybean oil emulsions, octane-in-water emulsions, and fat emulsions. SdFFF is also able to monitor changes in emulsion caused by aging or by the addition of electrolytes. SdFFF has been used to sort liposomes, as unilamellar vesicles or much larger multilamellar vesicles, the cubosom, and polylactate nanoparticles used as drug delivery systems [41]. 

Most of the above membrane-oriented studies were carried out for peptides in multilayer systems that were collapsed or transferred onto a sample cell surface. An alternative and very interesting way to study membrane systems is by IRRAS (infrared reflection absorption spectroscopy) at the air-water interface. In this way, unilamellar systems can be studied as a function of surface pressure and under the influence of various membrane proteins and peptides added. Mendelsohn et al.[136] have studied a model series of peptides, [K2(LA) ] (n = 6, 8, 10, 12), in nonaqueous (solution), multilamellar (lipid), and unilamellar (peptide-IRRAS) conditions. In the multilamellar vesicles these peptides are predominantly helical in conformation, but as peptide only monolayers on a D20 subphase the conformation is (1-sheet like, at least initially. For different lengths, the peptides show variable surface pressure sensitivity to development of some helical component. These authors further use their IR data to hypothesize the existence of the less-usual parallel (i-sheet conformation in these peptides. A critical comparison is available for different secondary structures as detected using the IRRAS data for peptides on H20 and D20 subphasesJ137 ... 

Figure 9.21 Various types of vesicles/liposomes, the so-called small unilamellar vesicles, SUV the large unilamellar vesicles, LUV the multilamellar vesicles, MLV...

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